Monitoramento via PCR de Salmonella spp. no processamento de carne suína

Abstract

The Salmonella spp is major pathogens involved in Food borne Diseases (FBD), especially outbreaks involving the ingestion of meet and this products. The aim of this study was to evaluate the food safety in pork and sausages production for the Salmonella spp. presence in the process. The samples were collected in a refrigerator that slaughter pigs and manufactures sausages, located in the Campos Gerais - PR. Initially, to standardize the PCR was necessary to determine an extraction protocol, as well as methodological adjustments to the PCR reaction. For these tests it was used sample of Salmonella isolated from food. The extraction protocol was tested by heating process and for amplification reaction were evaluated three different concentration of DNA and hybridization temperatures to establish the ideal standard. The chosen protocol proved to be very efficient for the Salmonella ssp DNA extraction because it allowed obtaining DNA in sufficient quality and quantity for amplification bands. For amplification, the best condition was found a concentration of approximately 40 ng DNA and a hybridization temperature of 57 ° C. In order to validate the molecular analysis by PCR, we carried out a comparative study with the initial conventional microbiology for proof of all results obtained by molecular analysis. Seventeen points were chosen during the different stages of production process. Two carcasses were monitored throughout the procedure and samples were collected, comprising the scalding step to the final product inlay. The PCR usage technique proved advantageous in the following aspects: total analysis time of approximately 30 hours; higher sensitivity compared to conventional. After validation, we performed a new collection, covering stages from pre-slaughter until embedded obtaining, making a total of 62 samples, in order to assess contamination during production of pork and sausages. As a result, it was found that 60% of the samples were contaminated with Salmonella spp, in various stages of production. From this evaluation, we selected some points contaminated and developed a corrective action plan in order to control and reduce microbiological hazards on the premises. Through further analysis by PCR was possible to verify that the action plan was effective in 100% of samples.